Transformation of bacterial cells will be carried out using the plasmid _____. T

Question

Transformation of bacterial cells will be carried out using the plasmid _____. The bacteria ____ (Genus species) were grown overnight on bacterial plates In this transformation experiment, the host cells will be mixed with the transformation solution ____ to make them competent or ABLE to take up the plasmid DNA. Next, the bacterial cells will be mixed with the plasmid ______ over ice. This will be followed by a "heat shock" treatment at ___ degree C for a short period of _____ seconds that will drive the plasmid into the cells. The bacterial cells will then be allowed to "heal" or recover in ____ ______ at _____ or ____. If the bacterial cells successfully take up the plasmid DNA and the plasmid replicates within the bacteria (outside the genome) the cells are said to be ________. If the transformation is successful, the recombinant E. coli contain plasmid DNA which replicates within the cells. The plasmid DNA has coding regions (a.k.a. ____) for three proteins. These genes are called _____, ____, and _____. As the transformed bacteria grow and divide, the transformed cell becomes a colony and genes present on the plasmid are transcribed into _____ and then translated into _____. When a colony of transformed E. coli is plated on medium containing the antibiotic ampicillin expression of the _____ resistance gene allows it to grow. The antibiotic Ampicillin contains a ___ ring which is important for its inhibition of cell formation in E. coli. The protein coded for by this gene has the specific name _____ ____ because it destroys the Beta lactam ring. In this, the enzyme will be produced by the transformed bacteria that can grow in the presence of the substrate ______ that is added to the cultural medium. The transformed bacterial cells will then be plated onto two plates with medium the bacterial LB medium containing either ___ only or _______ + _____. Cells of colonies growing on the first plate produce two proteins: an enzyme called ______ and a regulatory protein called ____. Cells of colonies growing only on the second plate produce in addition the third protein __ which changes the colony color from _____ of the original untransformed cells to _____ color and to fluoresce when illuminated with UV.

Explanation / Answer

FIRST PARAGRAPH ANSWERS:

1. DNA

2. Escherichia coli

3. Calcium chloride(CaCl2)8

4. pUC 19

5. 48°C8

6. 30 seconds

7. LB medium

8. 37°C or 20°C

9. Daughter cells.

SECOND PARAGRAPH ANSWERS:

1. 5'-3'

2. transformation,transduction and conjugation.

3. DNA

4. RNA

5. ampicillin

6. Beta lactum ring

7. Proteome

8. LB broth.

THIRD PARAGRAPH ANSWERS:

1. LB nutrient agar only or LB nutrient agar+LB nutrient agar ampicillin.

2.Digestive enzyme

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